In vitro clonal propagation of biodiesel plant ( Jatropha curcas L ...

RESEARCH COMMUNICATIONS

In vitro clonal propagation of biodiesel
resources offering a clean substitute for expensive fossil
plant (Jatropha curcas L.)
fuel imports, thus enabling the country to meet the objectives
of economic growth, fuel security and cleaner air.

J.
curcas is the most primitive species of the genus and
Mukul Manjari Datta1, Priyanka Mukherjee1,
forms artificial and natural hybrid complexes readily and
Biswajit Ghosh2 and Timir Baran Jha1,*
possess a problem to the genetic fidelity1. Conventional agri-
1
culture uses seeds and cuttings for its propagation. But
Department of Botany, Presidency College, Kolkata 700 073, India
2Department of Botany, RKMVC College, Rahara, Kolkata 700 118, India
the seeds are heterozygous in nature and the cuttings are

seasonal. Moreover, it is reported that vegetative cuttings
In the present investigation, in vitro clonal propagation
are not deep-rooted and are easily uprooted as they do not
of seven-month-old Jatropha curcas L. was achieved
form a taproot system2. Seed set has been reported to be low
employing nodal explants. Axillary shoot bud prolifera-
in vegetatively propagated plants2.
tion was best initiated on Murashige and Skoog’s (MS)
Tissue culture studies were undertaken in different spe-
basal medium supplemented with 22.2 μM N6-benzyl-
cies of Jatropha. Morphogenesis from endosperm tissues
adenine (BA) and 55.6 μM adenine sulphate, in which
has been reported in J. panduraefolia3–5. High frequency
cultures produced 6.2 ± 0.56 shoots per nodal explant
regeneration from various explants of J. integerrima has
with 2.0 ± 0.18 cm average length after 4–6 weeks. The
rate of shoot multiplication was significantly enhanced
been reported6. Using different explants, plant regenera-
after transfer to MS basal medium supplemented with
tion protocols have also been described in J. curcas2,7–9,
2.3 μM 6-furfuryl amino purine (Kn), 0.5 μM indole-
but multiplication rate was low for field applications. More-
3-butyric acid (IBA) and 27.8 μM adenine sulphate for
over, no lab-to-land transfer protocol of J. curcas using
4 weeks. Both shoot number (30.8 ± 5.48) and average
nodal meristems is available. Nodal meristems are an im-
shoot length (4.8 ± 0.43 cm) were found to increase
portant source tissue of micropropagation and plants
significantly. About 52% of root induction occurred in
raised from these are comparatively more resistant to gene-
MS basal medium supplemented with 1.0 μM IBA in
tic variation10. Keeping in mind the economical impor-
2–3 weeks. Further elongation of roots with average
tance of J. curcas, critical analysis of the earlier protocols
length of 8.7 ± 1.35 cm was obtained in unsupplemented
necessitated formulating a well-documented, reproducible,
MS basal medium for 2–3 weeks. The plantlets (12–16-
in vitro micropropagation protocol.
week-old) were successfully acclimatized in soil with
87% survival frequency.
In the present communication, we report an efficient

regeneration system of micropropagation from nodal ex-
Keywords: Axillary shoot, clonal propagation, Jatropha
plants of a seven-month-old J. curcas plant and establish-
curcas, nodal explants.
ment of plants under field conditions.

J. curcas L. seeds collected from Ramakrishna Mis-
J
sion, Narendrapur, West Bengal were sown in the experi-
ATROPHA CURCAS L. (Euphorbiaceae) or physic nut is an
all-purpose, zero-waste perennial plant. It is considered as
mental garden of Presidency College, Kolkata. The seedlings
a potential source of non-edible fuel-producing plant along
were grown in sterile vermiculite at 25–30°C in light. All
with its different medicinal properties and grows well in
the explants were collected from this donor plant for the
the tropical and subtropical climate in India. The seeds
present investigation.
contain 4–40% viscous oil known as ‘curcas oil’. The oil
Nodal explants (2–3 cm in length) collected from the
is high in cetane value and can be used directly in diesel
seven-month-old donor plant were kept for 3 h in a systemic
engines added to diesel fuel as an extender or trans-
fungicide, Bavistin (BASF India Ltd) prior to surface-
esterized to a biodiesel fuel. The oil is a clean fuel reducing
sterilization. They were surface-sterilized in 0.1% HgCl2
greenhouse gas emissions, has greater lubricity and re-
(w/v) for 20–25 min followed by repeated washing (five
duces engine wear. Pure Jatropha biodiesel is non-toxic
times) with sterile distilled water. After sterilization, the
in nature. This oil is a strong purgative, widely used as an
explants were trimmed (~1.0 cm) at the base and cultured
antiseptic for cough, skin diseases and as a pain reliever
with the cut surface in contact with the culture medium
in rheumatism. Refining crude Jatropha oil into bio-fuel
(Figure 1 a).
products produces glycerine as by-product, which is in
For axillary shoot bud proliferation, the nodal explants
great demand as a raw material for cosmetic, medicine
cultured on Murashige and Skoog (MS)11 basal medium
and food product industries.
at full and half strength, supplemented with 3% (w/v) su-
India has growing energy and transport fuel demand,
crose were studied. MS basal medium was supplemented
where J. curcas has the potential to become one of the
with 55.6 μM adenine sulphate; different cytokinins, e.g.
world’s key energy crops. However, inexpensive biodiesel
2.3–37.2 μM 6-furfurylamino purine (Kn), 2.2–35.6 μM
can be produced from India’s vast agribiotechnological
N6-benzyladenine (BA), 2.3–36.4 μM (thidiazuron; TDZ)
or 2.5–39.4 μM 2-iP (2-isopentenyladenine) individually
for 4–6 weeks (Sigma, Aldrich, USA). All media were so-
*For correspondence. (e-mail: presibot@vsnl.net)
lidified with 0.75% (w/v) agar (Merck, India).
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RESEARCH COMMUNICATIONS

All experiments were carried out in culture tubes (150 ×
supplemented with 3% (w/v) sucrose and different con-
25 mm) containing 20 ml of culture medium. The pH of
centrations of auxins were tested individually: 0.5–11.0 μM
all media was adjusted to 5.7 ± 0.1 prior to autoclaving at
α-napthaleneacetic acid (NAA); 0.5–10.0 μM IBA; 0.5–
121°C at 1.1 kg sq. cm for 15 min. Cultures were incubated
11.4 μM IAA (indoleacetic acid) for 2–3 weeks. The
under 16 h/8 h light/dark cycles (artificial light, 80 μmol
shoots were also tested on unsupplemented full or half
per sq. m per s) at 22 ± 2°C.
strength MS basal medium with 3% sucrose (w/v) for root
After 4–6 weeks of culture, the shoots cultured on MS
initiation. The rooted (10–13-week-old) shoots were then
basal medium supplemented with 3% (w/v) sucrose, 22.2–
transferred to MS basal medium supplemented with 3%
35.6 μM BA and 55.6 μM adenine sulphate were trans-
sucrose (w/v) for 2–3 weeks for further elongation. Addi-
ferred to MS basal medium supplemented with 3% (w/v)
tion of activated charcoal (0.5, 1.0%) to MS basal medium
sucrose, 27.8 μM adenine sulphate, 2.2–23.2 μM Kn and
supplemented with 3% sucrose (w/v) for root elongation
2.2–22.2 μM BA, singly or in combination, as well as in
was also tested.
combination with 0.5–5.0 μM IBA (indole butyric acid)
The complete rooted plantlets (12–16-week-old) were
for raising multiple shoots. The best response of (30.8 ±
hardened for three weeks in MS basal medium under dif-
5.48) shoots per node was obtained in medium containing
fuse light (16/8 h photoperiod) in culture room prior to
2.3 μM Kn, 27.8 μM adenine sulphate and 0.5 μM IBA.
field transfer. Then the plantlets were transferred to pots
For initiation of roots, the 8–10-week-old shoots (2.0–
containing a mixture of soil and vermiculite in the ratio
3.0 cm in length) were cultured on MS basal medium
1 : 1 and covered with polyethylene bags to maintain more

than 80% relative humidity, with temperature ranging

from 25°C to 30°C and kept in field conditions. About
87% of the plants survived after 3 weeks of hardening.
The experiments were set up in a completely randomized
design. Data were analysed by analysis of variance (ANOVA)
to detect significant differences between means12. Means
differing significantly were compared using Duncan’s
Multiple Range Test (DMRT) at 5% probability level.
Variability of data was expressed as mean ± standard
error (SE).
Initiation of cultures from the nodal explants did not
pose a major problem. During initiation, the explants did
not show any leaching or browning of tissues. MS basal
medium was the most effective for in vitro shoot multi-
plication from nodal explants. The nodal explants cul-
tured on MS basal medium supplemented with 55.6 μM
adenine sulphate and different cytokinins showed varied
response (Table 1). Nodes cultured on half strength MS
basal medium showed no visible signs of tissue differen-
tiation. This was possibly due to a greater demand of nitrogen
and potassium-containing compounds, which induce greater
amount of new proteins13. These components are lower in
half strength MS basal medium compared to full strength
MS basal medium.
The best response (6.2 ± 0.56 shoots per nodal explant)
was obtained in the presence of 55.6 μM adenine sulphate,
22.2 μM BA and was found to be significantly (P ≤ 0.05
level) higher than shoots induced per nodal explant in
other concentrations of cytokinins (Kn, 2-iP or TDZ)
used in the present study (Table 1; Figure 1 b, c). How-
ever, at the same concentration of 22.2 μM BA, a lower


initiation (1.5 ± 0.2 shoot buds per explant; 2.2 ± 0.6
Figure 1. In vitro clonal propagation of biodiesel plant (Jatropha
shoots per nodal explant) of shoot bud proliferation
curcas L.). a, Nodal explants from the donor plant. b, Nodal explants
was reported2,8. This differential response may be attri-
after the first 2–3 weeks of culture. c, Axillary bud breaking after 4–6
weeks of culture. d, Profuse multiplication of shoots after 8–10 weeks
buted to the specific age and physiological condition of the
of culture. e, Initiation of roots in in vitro shoots. f, In vitro shoots with
donor plant from which the nodal explants were excised.
slender and white taproot system. g, Complete plantlet (12–16-week-
Nodes cultured on medium with different concentrations
old) grown under field conditions. h, Plant after 10 weeks of hardening.
i, Ten-month-old in vitro grown plant.
of Kn showed lower induction of axillary shoot-bud
CURRENT SCIENCE, VOL. 93, NO. 10, 25 NOVEMBER 2007
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RESEARCH COMMUNICATIONS

Table 1. Effect of different types and concentrations of cytokinins on shoot multiplication of Jatropha curcas
Media formulations:
MS basal medium + 3% sucrose (w/v)
Concentration of
Average number of shoots
Average length of shoots
+ 55.6 μM adenine sulphate +
cytokinin used (μM) (∼> 0.5 cm) developed per nodal explant
developed (cm ± SE)

Kinetin 2.3
2.2
± 0.17b 0.8
± 0.02a
4.6
2.0
± 0.10b 0.8
± 0.04a
11.6
2.4
± 0.12b 0.8
± 0.06a
23.2
3.5
± 0.35c 0.9
± 0.07a
37.2
1.0
± 0.01a 0.8
± 0.02a
BA 2.2
1.0
± 0.01a 0.8
± 0.02a
4.4
2.0
± 0.13b 0.8
± 0.05a
11.1
2.7
± 0.23bc 0.9
± 0.03a
22.2
6.2
± 0.56f 2.0
± 0.18b
35.6
5.3
± 0.40e 1.4
± 0.12ab
2-iP 2.5
1.0
± 0.03a 1.0
± 0.03a
4.9
1.2
± 0.05a 0.9
± 0.02a
12.3
1.0
± 0.02a 0.8
± 0.02a
24.6
1.0
± 0.01a 0.9
± 0.05a
39.4
1.0
± 0.03a 0.8
± 0.04a
TDZ 2.3
3.0
± 0.02c 0.8
± 0.03a
4.5
1.2
± 0.04a 0.9
± 0.06a
11.4
1.0
± 0.05a 0.9
± 0.02a
22.7
1.0
± 0.02a 0.8
± 0.03a
36.4
1.0
± 0.01a 0.9
± 0.02a
Data were recorded after 4–6 weeks of culture. Each treatment was repeated thrice and each replicate consisted of 3–5 nodal explants.
Means having different letters as superscripts are significantly different from each other (P ≤ 0.05) according to DMRT.


proliferation. Though TDZ is known to induce cytokinin-
was possibly due to a combined effect of different growth
like effects in a number of plant species, particularly regulators along with other additives. Lowering of growth
woody14 as well as herbaceous crop species15, the present
regulators in micropropagation studies to achieve higher
study showed negligible effects of TDZ on induction of
rate of multiplication has been reported in Holarrhena
shoot multiplication as also reported in J. curcas earlier2.
antidysenterica18. However, it has been observed in Jat-
Jatropha belongs to the family Euphorbiaceae and our
ropha that it requires higher concentration of only one
observation confirms earlier reports16,17 that among the type of cytokinin (BA) for induction phase and favours
cytokinins BA plays an important role in initiation of
lower concentration of another type of cytokinin (Kn)
shoot-bud proliferation in many members of the Euphor-
along with other additives for escalation and proliferation
biaceae family.
of shoot cultures. A recent report2 indicated similar effect
The 4–6-week-old-nodal cultures on MS basal medium
in J. curcas with BA and TDZ, but the authors obtained
supplemented with 55.6 μM adenine sulphate and 22.2–
lesser number of shoots (2.0 ± 0.8 to 12.3 ± 1.7) with
35.6 μM BA on transfer to MS basal medium supple-
comparatively higher amount of cytokinin.
mented with 27.8 μM adenine sulphate with cytokinin No roots could be induced in either MS or half strength
singly or in combination, as well as in combination with
MS basal medium, but profuse callusing at the base of the
IBA showed varied response (Table 2). In the present
shoots was noted in the presence of NAA and IAA. How-
study, transfer of cultures to MS basal medium with ever, when 2.0–3.0 cm elongated shoots were placed on MS
lower concentration of adenine sulphate (27.8 μM) and with
medium with lower concentration of 0.5–1.0 μM IBA,
same (22.2 μM) or reduced (2.2–11.1 μM) concentration
roots were induced in 52% of the shoots within three
of BA, led to induction of callus at the base and failed to
weeks (Table 3; Figure 1 e). Other concentrations of 2.5–
induce further shoot proliferation (Table 2). Results also
10.0 μM IBA did not induce any roots. A distinct taproot
indicated that shoots cultured on medium with combinations
system developed with slender and white secondary roots;
of BA and Kn did not proliferate further. On the other
this was considered important for hardening and field
hand, transferring the cultures from a set having higher
transfer (Figure 1 f ). Significant increase in root length
concentration of 22.2 μM BA and 55.6 μM adenine sul-
occurred on transfer to only MS basal medium without
phate to a set with lower concentration of 2.3 μM Kn,
addition of charcoal8 for another 2–3 weeks. It is to be
27.8 μM adenine sulphate and 0.5 μM IBA, led to a sig-
mentioned in this regard that rooting was obtained in MS
nificantly higher nodal shoot multiplication (30.8 ± 5.48
basal medium supplemented2 with 5.4 μM NAA, and in
shoots per nodal explant) within the next 4 weeks, which
MS basal medium supplemented8 with higher amount of
was not recorded previously (Table 2; Figure 1 d). This
15.0 μM IBA.
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Table 2. Effect of growth regulators on shoot multiplication from nodal explants of J. curcas

Growth regulators (μM)*

BA
IBA
Kinetin
Number of shoots per explant
Length of shoots

2.2 0
0
1.0
± 0.01a 0.8
± 0.02a
4.4 0
0
2.0
± 0.13b 0.8
± 0.05a
11.1 0
0
3.0
± 0.25a
1.0 ± 0.05a
22.2 0
0
6.5
± 0.60c 2.2
± 0.19b
2.2 0
2.3
1.0
± 0.02a 0.9
± 0.06a
4.4 0
4.6
1.2
± 0.05a 0.8
± 0.05a
11.1 0
11.6
1.0
± 0.03a 0.9
± 0.08a
22.2 0
23.2
1.0
± 0.01a 0.8
± 0.04a
2.2 0.5 0
1.2
± 0.08a 1.0
± 0.05a
4.4 1.0 0
1.5
± 0.04a 0.8
± 0.06a
11.1 2.5
0
1.3
± 0.03a 0.9
± 0.03a
22.2 5.0
0
1.0
± 0.02 0.9
± 0.01a
0 0.5 2.3
30.8
± 5.48g 4.8
± 0.43d
0 1.0 4.6
14.6
± 2.93f 3.9
± 0.48c
0 2.5
11.6
11.2
± 2.73e 3.6
± 0.32c
0 5.0
23.2
9.8
± 1.86d 3.0
± 0.29c
0 0 2.3
2.4
± 0.25a 0.9
± 0.04a
0 0 4.6
2.2
± 0.22a 0.9
± 0.06a
0 0 11.6
2.6
± 0.28a 0.9
± 0.05a
0 0 23.2
3.7
± 0.36b 1.0
± 0.08a
*MS basal medium + 3% sucrose + adenine sulphate (27.8 μM) along with different plant growth regulators.
Data were recorded after 3–4 weeks of culture. Each treatment was repeated thrice and each replicate
consisted of 3–5 nodal explants. Means having different letters as superscripts are significantly different
from each other (P ≤ 0.05) according to DMRT.


Table 3. Effect of IBA on initiation of roots in excised shoots of J. curcas
meristems with much higher rate of multiplication. This
protocol can be used as a basic tool to commercialize cul-
IBA
Percentage of
Number of
Length of roots
(μM)*
root induction
roots per shoot
(cm ± SE)
tivation of the biodiesel plant.


0 0 0
0

0.5 15
1.0
± 0.06a 1.2
± 0.09a
1. Prabakaran, A. J. and Sujatha, M., Jatropha tanjorensis Ellis &
1.0 52
5.6
± 0.42b
8.7 ± 1.35b
Saroja, a natural interspecific hybrid occurring in Tamil Nadu, In-
2.5 0 0
0
dia. Genet. Resour. Crop Evol., 1999, 46, 213–218.
5.0 0 0
0
2. Sujatha, M., Makkar, H. P. S. and Becker, K., Shoot bud prolifera-
10.0 0
0
0
tion from axillary nodes and leaf sections of non-toxic Jatropha
curcas L. Plant Growth Reg., 2006, 47, 83–90.
*MS basal medium + 3% sucrose.
3. Srivastava, P. S., In vitro induction of triploid roots and shoots
Data were recorded after 2–3 weeks of culture. Each treatment was re-
from mature endosperm of Jatropha panduraefolia. Z. Pflanzen-
peated thrice and each replicate consisted of 3–5 shoots. Means having
physiol., 1971, 66, 93–96.
different letters as superscripts are significantly different from each
4. Johri, B. M. and Srivastava, P. S., Morphogenesis in endosperm
other (P ≤ 0.05) according to DMRT.
cultures. Z. Pflanzenphysiol., 1973, 70, 285–304.

5. Srivastava, P. S. and Johri, B. M., Morphogenesis in mature endo-

sperm cultures of Jatropha panduraefolia. Beitr. Biol. Planz.,
1974, 50, 255–268.
The shoots (12–16-week-old) were removed from the
6. Sujatha, M. and Dhingra, M., Rapid plant regeneration from vari-
medium, thoroughly washed with water, dipped for 1 h in
ous explants of Jatropha integerrima. Plant Cell Tissue Organ
0.1% (w/v) bavistin (systemic fungicide), and trans-
Cult., 1993, 35, 293–296.
planted to plastic pots containing a mixture of (1 : 1) soil
7. Qin, W., Wei-Da, L., Yi, L., Shu-Lin, P., Ying, X. U., Lin, T. and
and vermiculite (Figure 1 f ). The plantlets were irrigated
Fang, C., Plant regeneration from epicotyl explants of Jatropha
curcas. J. Plant Physiol. Mol. Biol., 2004, 30, 475–478.
with tap water as and when required. Polythene covers
8. Rajore, S. and Batra, A., Efficient plant regeneration via shoot tip
were completely withdrawn after 3–5 weeks of hardening
explant in Jatropha curcas. J. Plant Biochem. Biotechnol., 2005,
(Figure 1 g). Plants were then transferred to potted soil
14, 73–75.
for further growth (Figure 1 h). The plants ranged from
9. Sujatha, M. and Mukta, N., Morphogenesis and plant regeneration
86.0 ± 0.12 to 102.0 ± 0.21 cm in height after 10 months
from tissue cultures of Jatropha curcas. Plant Cell Tissue Organ
Cult., 1996, 44, 135–141.
(Figure 1i).
10. Pierik, R. L. M., Commercial aspects of micropropagation. In
The present study describes a well-documented and re-
Horticulture – New Technologies and Applications (eds Prakash,
liable micropropagation protocol of J. curcas from nodal
J. K. and Pierik, R. L. M.), Dordercht, The Netherlands, 1991.
CURRENT SCIENCE, VOL. 93, NO. 10, 25 NOVEMBER 2007
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11. Murashige, T. and Skoog, F., A revised medium for rapid growth
work2. We recently collected three specimens resembling
and bioassays with tobacco tissue cultures. Physiol. Plant., 1962,
each other which fit the generic diagnosis given by Ravi-
15, 473–479.
chandran et al.3 for Gegeneophis, but which differ from
12. Sokal, R. and Rohlf, F. J., Introduction to Biostatistics, W.H.
Freeman, NY, 1987, 2nd edn.
all known species. Here we describe this form as a new
13. Guru, S. K., Chandra, R., Khetrapal, S., Raj, A. and Palisetty, R.,
species.
Protein pattern in differentiating explants of chick pea (Cicer
Gegeneophis mhadeiensis sp. nov. Figures 1–3. Holo-
arietinum L.). J. Plant Physiol., 1999, 4, 147–151.
type: Bombay Natural History Society, Mumbai, India
14. Barrueto Cid, L. P., Machado, A. C. M. G., Carvalheira, S. B. R.
(BNHS/4643). A mature male, collected from the sur-
C. and Brasileiro, A. C. M., Plant regeneration from seedling ex-
plants of Eucalyptus grandis × Europhylla. Plant Cell Tissue roundings of Rameshwar temple, 15°39′ N lat., 74°08′E
Organ Cult., 1999, 56, 17–23.
long. (Chorla Village, Khanapur Taluk, Belgaum District,
15. Huetteman, C. A. and Preece, J. E., Thidiazuron: A potent cyto-
Karnataka), July 2006. The locality is situated at 728 msl,
kinin for woody plant tissue culture. Plant Cell Tissue Organ
adjacent to the Mahadayi Wildlife Sanctuary in the West-
Cult., 1993, 33, 105–109.
ern Ghats region.
16. Ripley, K. P. and Preece, J. E., Micropropagation of Euphorbia
lathyrus L. Plant Cell Tissue Organ Cult., 1986, 5, 213–218.
Paratypes: Zoological Survey of India, Calicut (ZSI/
17. Tiedman, J. and Hawker, J. S., In vitro propagation of latex pro-
WGFRS/V/A/640), an immature female, collected from
ducing plants. Ann. Bot., 1982, 49, 273–279.
Chavatyaar, Chavato vaddo, Chorla village at 732 msl.
18. Kumar, R., Sharma, K. and Agrawal, V., In vitro clonal propaga-
This locality is about 2 km from the type locality and the
tion of Holarrhena antidysenterica (L.) (Wall.) through nodal ex-
other collection details are as for the holotype. BNHS/
plants of mature trees. In Vitro Cell Dev. Biol.–Plant, 2005, 41,
137–144.
4643, an albino, mature male collected from Chavatyaar;

other collection details are as for the holotype.
Received 29 January 2007; revised accepted 24 August 2007
Diagnosis:
A
species
of
Gegeneophis that fits the gene-
ric diagnosis of Ravichandran et al.3 differing from all other

species in the genus except G. carnosus, G. krishni and

G. nadkarnii in having more than 100 primary annuli

(117–122), but differing from G. carnosus (secondary
A new species of the Indian caecilian
grooves 7) and G. krishni (secondary grooves 13–15) in
genus Gegeneophis Peters (Amphibia:
having far more (29–31) primary annuli with secondary
Gymnophiona: Caeciliidae) from the
annular grooves, and from G. nadkarnii (secondary grooves
86) in having lower number of secondary annular grooves
surroundings of Mahadayi Wildlife
(29–31).
Sanctuary, Western Ghats
Description of holotype: Some morphometric and meri-

stic data are given in Table 1. The specimen is in good
condition generally, except for minor artifacts associated
Gopalakrishna Bhatta1,*, K. P. Dinesh2,
with preservation; notably mid-ventral longitudinal groove
P. Prashanth3 and Nirmal U. Kulkarni4
of 134 mm length extends between the third nuchal
1Department of Biology, BASE, Basavanagudi, Bangalore 560 004, India
groove and the vent. There is a 20 mm long mid-ventral
2Zoological Survey of India, Western Ghats Field Research Station,
incision into the body cavity beginning 41 mm in front of
Annie Hall Road, Calicut 673 002, India
3Agumbe Rainforest Research Station, Agumbe 577 411, India
the vent. There are several small scratches on the skin on
4Hiru Naik Bldg, Dhuler Mapusa 403 507, India
both the dorsal and ventral surfaces made during the

search for scales.
A new species of Indian caecilian, Gegeneophis mhade-
The body in life is sub-cylindrical and slightly dorso-
iensis (Amphibia: Gymnophiona: Caeciliidae) is des-
ventrally compressed (Figure 1), though almost uniform
cribed based on three specimens collected from the
in its width throughout (Table 1). In dorsal view, the head
Western Ghats, northern Karnataka, India. The spe-
tapers strongly from the level of the occiput to the tenta-
cies is distinguished from all other Gegeneophis by a
cular apertures. Anteriorly, the head tapers and terminates
combination of the number of primary and secondary
in a bluntly rounded but narrow snout tip. The posterior
annuli. One of the paratypes is an albino.
part of the head is slightly narrower than the nuchal re-

gion. In lateral view, the top of the head is straight and
Keywords: Caecilians,
Gegeneophis mhadeiensis, Maha-
without any strong bulges. The margin of the upper lip is
dayi Wildlife Sanctuary.
slightly arched. The distance between the jaw angle and

the top of the head is less than the distance between the jaw
THE Indian caecilian (Gymnophiona) genus Gegeneophis
angle and the ventral surface of the lower jaw. In ventral
contains nine nominate species1, with six of these having
view, the anterior margin of the lower jaw is more broadly
been described since 1999 in a wave of new systematic
rounded than the anterior margin of the snout.
The small sub-circular nostrils are close to the front of
*For correspondence. (e-mail: gkb@sancharnet.in)
the snout tip, and are visible dorsally and laterally but not
1442
CURRENT SCIENCE, VOL. 93, NO. 10, 25 NOVEMBER 2007

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